culture of Ophiostoma novo-ulmi

Paul Zambino paulz at PUCCINI.CRL.UMN.EDU
Fri Aug 2 09:44:39 EST 1996

>Dear all
>Could anyone please share with me some information on the most
>successful culture method for the Dutch Elm Disease fungus.
>Dr Stephan Helfer, SSO
>Mycologist / Plant Pathologist
>Royal Botanic Garden Edinburgh, Inverleith Row, EDINBURGH EH3 5LR,
>Scotland UK
>phone: +44 (0)131 552 7171 ext 280
>    or +44 (0)131 459 0446-280 (direct digital VoiceMail line)
>fax:   +44 (0)131 552 0382

Ophiostoma ulmi, and other species of Ophiostoma previously included in
Ceratocystis, related Europhium species, and anamorphs of Ophiostoma in
Sporothrix, Leptographium, Graphium  can all be semi-selectively isolated
using solid media that contains cycloheximide.  See Harrington, T. C.,
1981.  Cycloheximide sensitivity as a taxonomic character in Ceratocystis.
Mycologia 73:1123-1129.  For isolation, 1 to 1.5 % malt extract .1% yeast
extract, 1.5 % agar and 200 ppm cycloheximide added after autoclaving.
After isolation, delete the cycloheximide.

The cycloheximide will probably be unneccessary if isolating from surface
disinfested twig sections, but is very useful for isolating from dead bark
beetles.  Note that Beauveria frequently colonizes insect cadavers and can
also grow on this cycloheximide medium, and cycloheximide has been used in
a selective medium for Beauveria and Metarrhizium (Veen and Ferron. 1966.
A Selective medium for the isolation of Beauveria tenella and of
Metarrizium anisopliae.  J. Inv. Pathol. 8:168-9).

Good luck, Paul Zambino

Paul Zambino, Ph.D.
USDA Forest Service
Forestry Sciences Lab
5985 Highway K
Rhinelander, WI 54501
FAX: (715)362-1166
EMAIL: paulz at

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